中山大学徐亮课题通过RNA自组装控制的CRISPR/Cas9功能建立内源性基因连接。2021年11月17日，国际知名学术期刊《美国化学会杂志》在线发表了这一成果。

研究人员提出了一种自组装诱导的RNA回路，可以直接在内源性基因之间建立调节连接。受CRISPR/Cas9复合体中向导RNA的自然组装过程启发，这种设计采用了独立的触发RNA链来诱导形成三元引导RNA组装，以实现对CRISPR/Cas9的功能控制。通过这一总体原则，内源性基因的表达调控可以通过可激活的CRISPR/Cas9功能在大肠杆菌中由完全独立的内源性小RNA和mRNA控制。

此外，大肠杆菌的细胞表型随着新基因连接的引入而成功地被编程。此外，这一设计的功能也在哺乳动物系统中得到了验证。这种基于自组装的RNA回路表现出极大的灵活性和设计的简单性，为建立内源性基因连接提供了一种独特的方法，从而为操纵细胞遗传网络铺平了一条广阔的道路。

据介绍，构建能够人为地建立内源性基因连接的合成回路对于引入新的细胞行为表型至关重要。鉴于内源基因的多样性，它缺乏一个通用的、易于设计的工具箱来操纵基因网络。

附：英文原文

Title: Building Endogenous Gene Connections through RNA Self-Assembly Controlled CRISPR/Cas9 Function

Author: Jiao Lin, Wei-Jia Wang, Yang Wang, Yan Liu, Liang Xu

Issue&Volume: November 17, 2021

Abstract: Construction of synthetic circuits that can artificially establish endogenous gene connections is essential to introduce new phenotypes for cellular behaviors. Given the diversity of endogenous genes, it lacks a general and easy-to-design toolbox to manipulate the genetic network. Here we present a type of self-assembly-induced RNA circuit that can directly build regulatory connections between endogenous genes. Inspired from the natural assembling process of guide RNA in the CRISPR/Cas9 complex, this design employs an independent trigger RNA strand to induce the formation of a ternary guide RNA assembly for functional control of CRISPR/Cas9. With this general principle, expressional regulations of endogenous genes can be controlled by totally independent endogenous small RNAs and mRNAs in E. coli via activatable CRISPR/Cas9 function. Moreover, the cellular phenotype of E. coli is successfully programmed with introduction of new gene connections. In addition, the functionality of this design is also verified in the mammalian system. This self-assembly-based RNA circuit exhibits a great flexibility and simplicity of design and provides a unique approach to build endogenous gene connections, which paves a broad way toward manipulation of cellular genetic networks.

DOI: 10.1021/jacs.1c09041

Source: https://pubs.acs.org/doi/10.1021/jacs.1c09041